CN116440078A - Multi-channel Ca for promoting tumor calcium overload 2+ Preparation method of nano regulator - Google Patents
Multi-channel Ca for promoting tumor calcium overload 2+ Preparation method of nano regulator Download PDFInfo
- Publication number
- CN116440078A CN116440078A CN202310036155.XA CN202310036155A CN116440078A CN 116440078 A CN116440078 A CN 116440078A CN 202310036155 A CN202310036155 A CN 202310036155A CN 116440078 A CN116440078 A CN 116440078A
- Authority
- CN
- China
- Prior art keywords
- cao
- preparation
- parg
- nano
- calcium
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 239000011575 calcium Substances 0.000 title claims abstract description 42
- 238000002360 preparation method Methods 0.000 title claims abstract description 33
- 230000001964 calcium overload Effects 0.000 title claims abstract description 31
- 206010028980 Neoplasm Diseases 0.000 title claims abstract description 26
- 230000001737 promoting effect Effects 0.000 title claims abstract description 16
- 239000002105 nanoparticle Substances 0.000 claims abstract description 67
- 101150003479 Parg gene Proteins 0.000 claims abstract description 54
- 239000002502 liposome Substances 0.000 claims abstract description 29
- 238000011282 treatment Methods 0.000 claims abstract description 15
- 239000003814 drug Substances 0.000 claims abstract description 13
- 239000002245 particle Substances 0.000 claims abstract description 13
- 230000000259 anti-tumor effect Effects 0.000 claims abstract description 12
- 238000000034 method Methods 0.000 claims abstract description 11
- 229940079593 drug Drugs 0.000 claims abstract description 8
- 230000008569 process Effects 0.000 claims abstract description 8
- 230000003647 oxidation Effects 0.000 claims abstract description 6
- 238000007254 oxidation reaction Methods 0.000 claims abstract description 6
- 238000001556 precipitation Methods 0.000 claims abstract description 5
- 230000002194 synthesizing effect Effects 0.000 claims abstract description 4
- XEEYBQQBJWHFJM-UHFFFAOYSA-N iron Substances [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 claims description 107
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 87
- 239000000243 solution Substances 0.000 claims description 39
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 36
- 239000007864 aqueous solution Substances 0.000 claims description 18
- 238000005406 washing Methods 0.000 claims description 17
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 claims description 14
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 14
- 210000002472 endoplasmic reticulum Anatomy 0.000 claims description 12
- 239000000725 suspension Substances 0.000 claims description 11
- 229920000724 poly(L-arginine) polymer Polymers 0.000 claims description 10
- 238000003756 stirring Methods 0.000 claims description 10
- 229910021642 ultra pure water Inorganic materials 0.000 claims description 9
- 239000012498 ultrapure water Substances 0.000 claims description 9
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 8
- 230000002209 hydrophobic effect Effects 0.000 claims description 8
- 239000004343 Calcium peroxide Substances 0.000 claims description 7
- LHJQIRIGXXHNLA-UHFFFAOYSA-N calcium peroxide Chemical compound [Ca+2].[O-][O-] LHJQIRIGXXHNLA-UHFFFAOYSA-N 0.000 claims description 7
- 235000019402 calcium peroxide Nutrition 0.000 claims description 7
- 235000012000 cholesterol Nutrition 0.000 claims description 7
- 230000000694 effects Effects 0.000 claims description 7
- 229910052742 iron Inorganic materials 0.000 claims description 7
- 150000002632 lipids Chemical class 0.000 claims description 7
- 108010011110 polyarginine Proteins 0.000 claims description 7
- 239000002994 raw material Substances 0.000 claims description 7
- IIZPXYDJLKNOIY-JXPKJXOSSA-N 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCC\C=C/C\C=C/C\C=C/C\C=C/CCCCC IIZPXYDJLKNOIY-JXPKJXOSSA-N 0.000 claims description 6
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 claims description 6
- 229920002385 Sodium hyaluronate Polymers 0.000 claims description 6
- 238000005119 centrifugation Methods 0.000 claims description 6
- 239000011248 coating agent Substances 0.000 claims description 6
- 238000000576 coating method Methods 0.000 claims description 6
- 239000011246 composite particle Substances 0.000 claims description 6
- 239000000787 lecithin Substances 0.000 claims description 6
- 235000010445 lecithin Nutrition 0.000 claims description 6
- 229940067606 lecithin Drugs 0.000 claims description 6
- 229940010747 sodium hyaluronate Drugs 0.000 claims description 6
- YWIVKILSMZOHHF-QJZPQSOGSA-N sodium;(2s,3s,4s,5r,6r)-6-[(2s,3r,4r,5s,6r)-3-acetamido-2-[(2s,3s,4r,5r,6r)-6-[(2r,3r,4r,5s,6r)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2- Chemical compound [Na+].CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 YWIVKILSMZOHHF-QJZPQSOGSA-N 0.000 claims description 6
- 239000002904 solvent Substances 0.000 claims description 6
- 239000000126 substance Substances 0.000 claims description 6
- 238000002560 therapeutic procedure Methods 0.000 claims description 6
- 238000006243 chemical reaction Methods 0.000 claims description 5
- 230000036571 hydration Effects 0.000 claims description 5
- 238000006703 hydration reaction Methods 0.000 claims description 5
- 150000002500 ions Chemical class 0.000 claims description 5
- 238000002390 rotary evaporation Methods 0.000 claims description 5
- JQWAHKMIYCERGA-UHFFFAOYSA-N (2-nonanoyloxy-3-octadeca-9,12-dienoyloxypropoxy)-[2-(trimethylazaniumyl)ethyl]phosphinate Chemical group CCCCCCCCC(=O)OC(COP([O-])(=O)CC[N+](C)(C)C)COC(=O)CCCCCCCC=CCC=CCCCCC JQWAHKMIYCERGA-UHFFFAOYSA-N 0.000 claims description 4
- 239000000872 buffer Substances 0.000 claims description 4
- 238000004108 freeze drying Methods 0.000 claims description 4
- 239000011521 glass Substances 0.000 claims description 4
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 claims description 3
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 claims description 3
- 239000002253 acid Substances 0.000 claims description 3
- 235000011114 ammonium hydroxide Nutrition 0.000 claims description 3
- 229910052791 calcium Inorganic materials 0.000 claims description 3
- 230000003197 catalytic effect Effects 0.000 claims description 3
- 239000003937 drug carrier Substances 0.000 claims description 3
- 238000001035 drying Methods 0.000 claims description 3
- 230000003993 interaction Effects 0.000 claims description 3
- 230000035484 reaction time Effects 0.000 claims description 3
- 238000001338 self-assembly Methods 0.000 claims description 3
- 238000002604 ultrasonography Methods 0.000 claims description 3
- 238000010009 beating Methods 0.000 claims description 2
- 238000011068 loading method Methods 0.000 claims description 2
- 231100000588 tumorigenic Toxicity 0.000 claims 9
- 230000000381 tumorigenic effect Effects 0.000 claims 9
- 210000004881 tumor cell Anatomy 0.000 abstract description 9
- 230000003834 intracellular effect Effects 0.000 abstract description 5
- 230000002438 mitochondrial effect Effects 0.000 abstract description 4
- 230000005775 apoptotic pathway Effects 0.000 abstract description 3
- 238000006555 catalytic reaction Methods 0.000 abstract description 3
- 239000012528 membrane Substances 0.000 abstract description 3
- 239000002246 antineoplastic agent Substances 0.000 abstract description 2
- 229940041181 antineoplastic drug Drugs 0.000 abstract description 2
- 210000004027 cell Anatomy 0.000 description 17
- MWUXSHHQAYIFBG-UHFFFAOYSA-N Nitric oxide Chemical compound O=[N] MWUXSHHQAYIFBG-UHFFFAOYSA-N 0.000 description 14
- 239000000047 product Substances 0.000 description 12
- 241000699670 Mus sp. Species 0.000 description 9
- 206010006187 Breast cancer Diseases 0.000 description 7
- 208000026310 Breast neoplasm Diseases 0.000 description 7
- 238000010586 diagram Methods 0.000 description 5
- WCDDVEOXEIYWFB-VXORFPGASA-N (2s,3s,4r,5r,6r)-3-[(2s,3r,5s,6r)-3-acetamido-5-hydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-4,5,6-trihydroxyoxane-2-carboxylic acid Chemical compound CC(=O)N[C@@H]1C[C@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](C(O)=O)O[C@@H](O)[C@H](O)[C@H]1O WCDDVEOXEIYWFB-VXORFPGASA-N 0.000 description 4
- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 description 4
- 230000006907 apoptotic process Effects 0.000 description 4
- 230000004094 calcium homeostasis Effects 0.000 description 4
- 229910001424 calcium ion Inorganic materials 0.000 description 4
- 238000012258 culturing Methods 0.000 description 4
- 229940014041 hyaluronate Drugs 0.000 description 4
- 238000009210 therapy by ultrasound Methods 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 3
- 238000004113 cell culture Methods 0.000 description 3
- 210000000805 cytoplasm Anatomy 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 230000001404 mediated effect Effects 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- ZKHQWZAMYRWXGA-KQYNXXCUSA-J ATP(4-) Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](COP([O-])(=O)OP([O-])(=O)OP([O-])([O-])=O)[C@@H](O)[C@H]1O ZKHQWZAMYRWXGA-KQYNXXCUSA-J 0.000 description 2
- ZKHQWZAMYRWXGA-UHFFFAOYSA-N Adenosine triphosphate Natural products C1=NC=2C(N)=NC=NC=2N1C1OC(COP(O)(=O)OP(O)(=O)OP(O)(O)=O)C(O)C1O ZKHQWZAMYRWXGA-UHFFFAOYSA-N 0.000 description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- 101710205660 Calcium-transporting ATPase Proteins 0.000 description 2
- 101710134161 Calcium-transporting ATPase sarcoplasmic/endoplasmic reticulum type Proteins 0.000 description 2
- 239000012148 binding buffer Substances 0.000 description 2
- 239000001506 calcium phosphate Substances 0.000 description 2
- 229910000389 calcium phosphate Inorganic materials 0.000 description 2
- 235000011010 calcium phosphates Nutrition 0.000 description 2
- 230000001086 cytosolic effect Effects 0.000 description 2
- 238000000684 flow cytometry Methods 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 230000003285 pharmacodynamic effect Effects 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 231100000048 toxicity data Toxicity 0.000 description 2
- 238000004627 transmission electron microscopy Methods 0.000 description 2
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 2
- 230000004614 tumor growth Effects 0.000 description 2
- RYGMFSIKBFXOCR-UHFFFAOYSA-N Copper Chemical compound [Cu] RYGMFSIKBFXOCR-UHFFFAOYSA-N 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 1
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 1
- 108010040476 FITC-annexin A5 Proteins 0.000 description 1
- 102000004310 Ion Channels Human genes 0.000 description 1
- ODKSFYDXXFIFQN-BYPYZUCNSA-N L-arginine Chemical compound OC(=O)[C@@H](N)CCCN=C(N)N ODKSFYDXXFIFQN-BYPYZUCNSA-N 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 108010019160 Pancreatin Proteins 0.000 description 1
- 206010039491 Sarcoma Diseases 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 230000003799 ca2+ efflux Effects 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 238000011254 conventional chemotherapy Methods 0.000 description 1
- 229910052802 copper Inorganic materials 0.000 description 1
- 239000010949 copper Substances 0.000 description 1
- 238000000354 decomposition reaction Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 230000008021 deposition Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000010494 dissociation reaction Methods 0.000 description 1
- 230000005593 dissociations Effects 0.000 description 1
- 238000002296 dynamic light scattering Methods 0.000 description 1
- 238000001493 electron microscopy Methods 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000029142 excretion Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 210000002744 extracellular matrix Anatomy 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000004941 influx Effects 0.000 description 1
- 238000010253 intravenous injection Methods 0.000 description 1
- 230000007794 irritation Effects 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 230000002132 lysosomal effect Effects 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000001000 micrograph Methods 0.000 description 1
- 208000012268 mitochondrial disease Diseases 0.000 description 1
- 210000001700 mitochondrial membrane Anatomy 0.000 description 1
- 239000002539 nanocarrier Substances 0.000 description 1
- 235000021590 normal diet Nutrition 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 230000036542 oxidative stress Effects 0.000 description 1
- 238000004806 packaging method and process Methods 0.000 description 1
- 229940055695 pancreatin Drugs 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 230000004526 pharmaceutical effect Effects 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 238000005464 sample preparation method Methods 0.000 description 1
- 230000011664 signaling Effects 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 231100000057 systemic toxicity Toxicity 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000026683 transduction Effects 0.000 description 1
- 238000010361 transduction Methods 0.000 description 1
- 210000001364 upper extremity Anatomy 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/10—Dispersions; Emulsions
- A61K9/127—Liposomes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/74—Synthetic polymeric materials
- A61K31/785—Polymers containing nitrogen
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K33/00—Medicinal preparations containing inorganic active ingredients
- A61K33/24—Heavy metals; Compounds thereof
- A61K33/26—Iron; Compounds thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K33/00—Medicinal preparations containing inorganic active ingredients
- A61K33/40—Peroxides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/24—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing atoms other than carbon, hydrogen, oxygen, halogen, nitrogen or sulfur, e.g. cyclomethicone or phospholipids
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Epidemiology (AREA)
- Inorganic Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Dispersion Chemistry (AREA)
- Biophysics (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Medicinal Preparation (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention provides a multi-channel Ca for promoting tumor calcium overload 2+ The preparation method of the nano regulator comprises the following steps: s1, synthesizing CaO through hydrolysis-precipitation process 2 A nanoparticle; s2, preparing CaO 2 ‑TA‑Fe 3+ A nanoparticle; s3, preparation of LipoCaO 2 ‑TA‑Fe 3+ (PArg) nanoparticles; when the medicine reaches the tumor part, the liposome membrane is broken and CaO is absorbed 2 ‑TA‑Fe 3+ Particles can generate OH and Ca 2+ Further oxidation of PArg by OH to NO, then OH, NO and self-supplied Ca by the nanoparticle 2+ By co-modulating endogenous and exogenous Ca in tumor cells 2+ Promoting intracellular calcium overload. So as to activate mitochondrial apoptosis pathway, destroy tumor cell skeleton and play the anti-tumor role of nano regulator; the preparation method is stable and reliable, the cost is low, and the prepared LipoCaO is 2 ‑TA‑Fe 3+ The PArg can play roles of cascade catalysis and ion interference in preparing anti-tumor drugs, and the multi-channel calcium overload promotion and anti-tumor effect enhancement are innovations in tumor treatment drugs.
Description
Technical Field
The invention relates to the field of biological medicine, in particular to a multi-channel Ca for promoting tumor calcium overload 2+ A method for preparing nano regulator.
Background
In recent years, anti-tumor therapies based on calcium overload have received increasing attention by disrupting mitochondrial ca2+ homeostasis, thereby overcoming the problems of drug-resistant O2 deficiency and poor biological safety encountered with conventional chemotherapy and PDT treatments. Under normal conditions of the cell, ca2+ is one of the most important second messengers, closely related to biosignal transduction, and is dedicated to regulating various physiological processes. However, when ca2+ homeostasis is disrupted, the abnormal calcium overload thus produced will involve a range of mitochondrial diseases including reduced Mitochondrial Membrane Potential (MMP), reduced Adenosine Triphosphate (ATP) levels and activation of mitochondrial-mediated apoptosis pathways. Therefore, ca2+ homeostasis by Ca2+ overload of bad tumor cells is expected to be one of the potential means of anti-tumor therapy.
To date, many researchers have focused on directly promoting ca2+ overload by delivering exogenous ca2+ to tumors. As shown in the previous report, tumor cells are more sensitive to ca2+ overload than normal cells due to remodelled ca2+ transport. Thus, several ca2+ nano-generators have been developed for use in calcium overload-mediated antitumor therapy, including calcium peroxide (CaO 2), calcium carbonate (CaCO 3), calcium phosphate (CaP). In addition, in order to ensure calcium overload and avoid induced systemic toxicity, various Tumor Microenvironment (TME) stimulus response designs have emerged for controlling ca2+ release in tumors, such as acidic pH, redox, enzymes, light, temperature, etc. Unfortunately, the antitumor effect of these ca2+ nano-generators is undesirable because intracellular ca2+ levels can quickly return to normal concentrations after rising due to the inherent ca2+ efflux channels.
Recently, another strategy for calcium overload has been to regulate endogenous ca2+ flow to the cytoplasm. Ca2+ is an exogenous messenger that is signaled in cells and is widely distributed in vivo. However, physiological calcium homeostasis maintains a huge ca2+ concentration gradient, with cytoplasmic ca2+ concentrations below 0.1 μm, while intracellular stored ca2+ is up to 0.28mM in the Endoplasmic Reticulum (ER) ca2+ reservoir, with extracellular ca2+ concentrations even above 1.5mM. Thus, modulation of endogenous ca2+ from the extracellular matrix and the Endoplasmic Reticulum (ER) to the cytoplasm will facilitate calcium overload in the cell. Recent studies indicate that OH mediated oxidative stress can raise intracellular ca2+ by opening the influx ion channel and shutting off the efflux calcium pump. Furthermore, the signaling molecule Nitric Oxide (NO) is reported to have the function of promoting calcium overload by opening RyR channels on the ER and promoting the storage of high concentrations of ca2+ from the Endoplasmic Reticulum (ER) (mM) to the cytoplasm (μm). However, the above single strategy is insufficient to achieve sufficient calcium overload to achieve effective anti-tumor treatment. Thus, the good synergy of Ca2+,. OH and NO produced in multichannel Ca2+ nano-modulators is used to amplify calcium overload, which has not been reported yet.
Disclosure of Invention
In order to make up for the defects, the invention provides a multi-channel Ca for promoting tumor calcium overload 2+ The preparation method of the nano regulator can effectively solve the problem of calcium ion excretion, realize cascade catalysis for treating tumors and ion interference for tumor cells, and promote apoptosis of the tumor cells.
The invention is realized in the following way:
multi-channel Ca for promoting tumor calcium overload 2+ The preparation method of the nano regulator comprises the following steps:
s1, synthesizing CaO through hydrolysis-precipitation process 2 Nanoparticles: adding CaCl into sodium hyaluronate solution 2 Adding ammonia water and 30% hydrogen peroxide solution in turn, adding sodium hydroxide aqueous solution under ultrasonic wave after stirring, washing with NaOH, water and ethanol, and freeze-drying for further use;
s2, preparing CaO 2 -TA-Fe 3+ Nanoparticles: caO prepared in S1 2 Dissolving the nano particles, and then sequentially adding TA aqueous solution and FeCl 3 Adding NaOH solution into the aqueous solution to adjust the pH value, centrifugally collecting the product, washing the product with ultrapure water for a plurality of times, and drying the product in a freeze dryer to obtain CaO 2 -TA-Fe 3+ A nanoparticle;
s3, preparing Lipo CaO 2 -TA-Fe 3+ (pag) nanoparticles: caO prepared in S2 2 -TA-Fe 3+ The nanoparticles were dissolved in ethanol, then dissolved in a glass flask together with the liposome raw material, film was formed by rotary evaporation, and then an aqueous solution of pagg was added to the flask under ultrasound, then sonicated at 0 ℃ for 3 minutes to obtain a uniform lipid suspension, the suspension was centrifuged for 10 minutes, washed three times with ultra-pure water to remove unencapsulated pagg, and finally, the liposomes were dispersed in PBS buffer.
In one embodiment of the present invention, the Lipo CaO in S3 2 -TA-Fe 3+ The PArg nanoparticle takes lecithin liposome as a drug carrier, and calcium peroxide composite particles and polyarginine as a hydrophobic layer and a hydrophilic cavity substance.
In one embodiment of the invention, the calcium peroxide composite particle is calcium peroxide-tannic acid-iron and has a chemical formula of CaO 2 -TA-Fe 3+ The polyarginine is poly-L-arginine, namely PArg.
In one embodiment of the present invention, the Lipo CaO 2 -TA-Fe 3+ The preparation of the/PArg nanoparticles is carried out by reacting CaO 2 -TA-Fe 3+ And lecithin, cholesterol, DSPE-PEG2000 in organic solution by rotary evaporation to form filmAnd then is obtained by the hydration of the PArg.
In one embodiment of the present invention, the sodium hyaluronate solvent in S1 is 10mL,15mg/mL, stirring at room temperature for more than 3h, and CaCl 2 The aqueous solution was 0.1g/mL, and NH 3 ·H 2 O is 1M, which is prepared in the prior art, and 30% hydrogen peroxide solution is H with the mass fraction of 30% 2 O 2 The stirring reaction time after the addition of the aqueous solution was 3 hours.
In one embodiment of the present invention, the aqueous NaOH solution in S1 is 0.1M, and the washing process of NaOH, water and ethanol is to first perform a washing treatment with NaOH to remove the excess acid, to wash with water to remove the neutralized solution, and then to wash with ethanol to remove the residual water stain.
In one embodiment of the invention, the TA concentration in S2 is 40mg/mL; fe (Fe) 3+ The concentration is 10mg/mL; TA and Fe 3+ The volume ratio of (1): 2, adding NaOH solution for adjusting the pH value, wherein the NaOH solution is 0.1M NaOH solution, and adjusting the pH value to 8.5; caO (CaO) 2 And TA, fe 3+ The reaction is carried out for 2 hours at room temperature; during centrifugation, the product was collected by rotation at 10000rpm for a centrifugation period of 10 min.
In one embodiment of the invention, the solvent in S3 is ethanol, chloroform, the liposome raw material component is egg yolk lecithin, the dosage is 50mg, the dosage of cholesterol is 9.6mg, the dosage of DSPE-PEG2000 is 6.37mg, and CaO 2 -TA-Fe 3+ The concentration of (2) is 10mg/mL, and the concentration of PArg is 15mg/mL; the hydration time of the aqueous PArg solution was 30min and the suspension was centrifuged at 10000rpm for 10min to collect the product.
In one embodiment of the present invention, the liposome in S3 encapsulates Lipo CaO 2 -TA-Fe 3+ In the preparation of nanoparticles of PArg, caO is first prepared 2 -TA-Fe 3+ Nanoparticle, synthetic CaO 2 Thereafter, TA-Fe is electrostatically reacted 3 + Self-assembly of coating on CaO 2 Form CaO on the surface of (C) 2 -TA-Fe 3+ Nanoparticles with a particle size of about 45nm, and then passing CaO through the liposomes 2 -TA-Fe 3+ The nano particles and the PArg are respectively wrapped in the hydrophobic layer and the hydrophilic cavity to obtain Lipo CaO 2 -TA-Fe 3+ (PArg) nanoparticles.
In one embodiment of the present invention, the Lipo CaO 2 -TA-Fe 3+ PArg is used to turn off Ca pump discharge by Fenton catalytic oxidation effect 2+ And opening Ca 2+ Internal flow channel and ion interference therapy to deliver exogenous Ca 2+ Gas NO treatment to release Ca in endoplasmic reticulum 2+ Application of the compound calcium-overload-promoting and tumor-treating medicine prepared by combining cytoplasmic triple striking interaction and CaO-loaded medicine 2 -TA-Fe 3+ And the liposomes of PArg, for use in the treatment of calcium overload and antitumor.
The beneficial effects of the invention are as follows:
CaO in the invention 2 -TA-Fe 3+ Nanoparticles trigger TA and Fe at lysosomal acidity 3+ Coating dissociation and CaO irritation 2 Decomposition of self-supplied Ca 2+ And H 2 O 2 . Wherein dissociated TA will drive Fe 3+ Reduction to Fe 2+ Followed by Fenton reaction with H 2 O 2 The reaction forms self-circulation high-oxidability OH, and the oxidation effect of OH can close Ca discharged from a calcium pump 2+ And opening Ca 2+ An inner flow channel to promote calcium overload;
more importantly, OH will oxidize PArg further to produce more NO. NO opens RyR channels in endoplasmic reticulum (mM) storage, releasing Ca in endoplasmic reticulum 2+ Further promoting calcium overload;
in addition, liposomes are the most common carriers in lipid-based nanocarriers, and are widely used for delivering drug molecules due to their unique ability to encapsulate hydrophilic and hydrophobic agents and targeting properties resulting from Enhanced Permeability and Retention (EPR) effects or ligand modification, and provide a platform for reducing tumor deposition and achieving drug release on demand, so pH-sensitive liposomes are used to encapsulate both drug molecules;
when the medicine reachesWhen the tumor is located, the liposome membrane is broken and CaO is contained 2 -TA-Fe 3+ Particles can generate OH and Ca 2+ Further oxidation of PArg by OH to NO, then OH, NO and self-supplied Ca by the nanoparticle 2+ By co-modulating endogenous and exogenous Ca in tumor cells 2+ Promoting intracellular calcium overload. So as to activate mitochondrial apoptosis pathway, destroy tumor cell skeleton and exert anti-tumor effect of nano regulator;
the preparation method of the invention is stable and reliable, and has low cost, and the prepared Lipo CaO 2 -TA-Fe 3+ The PArg can play roles of cascade catalysis and ion interference in preparing anti-tumor drugs, and the multi-channel calcium overload promotion and anti-tumor effect enhancement are innovations in tumor treatment drugs.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are needed in the embodiments will be briefly described below, it being understood that the following drawings only illustrate some examples of the present invention and therefore should not be considered as limiting the scope, and other related drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is CaO prepared in example 1 of the invention 2 -TA-Fe 3+ Schematic structural diagram of nanoparticle;
FIG. 2 is CaO prepared in example 1 of the present invention 2 -TA-Fe 3+ A transmission electron microscope image of the nanoparticle;
FIG. 3 is CaO prepared in example 1 of the invention 2 -TA-Fe 3+ Nanoparticles and CaO 2 Particle size and potential diagram of (a);
FIG. 4 is a diagram showing Lipo CaO prepared in example 2 of the present invention 2 -TA-Fe 3+ Transmission electron microscopy and particle size distribution of the/PArg nanoparticles;
FIG. 5 is a diagram of Lipo CaO prepared in example 2 of the present invention 2 -TA-Fe 3+ PArg nanoparticle and Lipo CaO2-TA-Fe 3+ 、Lipo CaO 2 Toxicity data graph for 4T1 cells;
FIG. 6 is the presentInventive example 2 Lipo CaO 2 -TA-Fe 3+ Schematic of the apoptosis of breast cancer cells 4T1 that/pagg can promote;
FIG. 7 is a diagram showing Lipo CaO prepared in example 2 of the present invention 2 -TA-Fe 3+ Experimental data graph of pharmacodynamic studies of the pag nanoparticles.
Detailed Description
For the purpose of making the objects, technical solutions and advantages of the embodiments of the present invention more apparent, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention, and it is apparent that the described embodiments are some embodiments of the present invention, but not all embodiments. All other embodiments, based on the embodiments of the invention, which are apparent to those of ordinary skill in the art without inventive faculty, are intended to be within the scope of the invention.
Referring to fig. 1-3, the present invention provides a technical solution: multi-channel Ca for promoting tumor calcium overload 2+ The preparation method of the nano regulator comprises the following steps:
s1, synthesizing CaO through hydrolysis-precipitation process 2 Nanoparticles: adding CaCl into sodium hyaluronate solution 2 Adding ammonia water and 30% hydrogen peroxide solution in turn, adding sodium hydroxide aqueous solution under ultrasonic wave after stirring, washing with NaOH, water and ethanol, and freeze-drying for further use;
s2, preparing CaO 2 -TA-Fe 3+ Nanoparticles: caO prepared in S1 2 Dissolving the nano particles, and then sequentially adding TA aqueous solution and FeCl 3 Adding NaOH solution into the aqueous solution to adjust the pH value, centrifugally collecting the product, washing the product with ultrapure water for a plurality of times, and drying the product in a freeze dryer to obtain CaO 2 -TA-Fe 3+ A nanoparticle;
s3, preparing Lipo CaO 2 -TA-Fe 3+ (pag) nanoparticles: caO prepared in S2 2 -TA-Fe 3+ Dissolving nanoparticles in ethanol, dissolving in glass flask together with liposome raw material, rotary evaporating to form film, and dissolving PArg in waterThe solution was added to the flask under ultrasound, then sonicated at 0 ℃ for 3 min to obtain a homogeneous lipid suspension, the suspension was centrifuged for 10min and washed three times with ultra pure water to remove unencapsulated pag, finally the liposomes were dispersed in PBS buffer.
To achieve CaO 2 -TA-Fe 3+ packaging/PArg to prepare nano-particles, in one embodiment of the invention, the Lipo CaO in S3 2 -TA-Fe 3+ The PArg nanoparticle takes lecithin liposome as a drug carrier, and calcium peroxide composite particles and polyarginine as a hydrophobic layer and a hydrophilic cavity substance.
In order to realize the arrangement of the hydrophobic layer and the hydrophilic cavity, caO is conveniently realized 2 -TA-Fe 3+ In one embodiment of the invention, the calcium peroxide composite particles are calcium peroxide-tannic acid-iron and have the chemical formula CaO 2 -TA-Fe 3+ The polyarginine is poly-L-arginine, namely PArg.
In order to achieve the preparation of liposomes and the formation of nanoparticles, in one embodiment of the invention, the Lipo CaO 2 -TA-Fe 3+ The preparation of the/PArg nanoparticles is carried out by reacting CaO 2 -TA-Fe 3+ And lecithin, cholesterol, DSPE-PEG2000 in organic solution, and then is hydrated by PArg.
To achieve CaO 2 Nanoparticle preparation in one embodiment of the present invention, the sodium Hyaluronate (HA) solvent in S1 is 10mL,15mg/mL, stirring at room temperature for more than 3h, and CaCl 2 The aqueous solution was 0.1g/mL, and NH 3 ·H 2 O is 1M, which is prepared in the prior art, and 30% hydrogen peroxide solution is H with the mass fraction of 30% 2 O 2 The stirring reaction time after the addition of the aqueous solution was 3 hours.
To achieve CaO 2 In one embodiment of the present invention, the aqueous solution of NaOH in S1 is 0.1M, and the washing process of NaOH, water and ethanol is to wash with NaOH to remove the excess acid,washing with water to remove the neutralized solution, and then washing with ethanol to remove residual water stains.
To achieve CaO 2 -TA-Fe 3+ Nanoparticle preparation, in one embodiment of the invention, the TA concentration in S2 is 40mg/mL; fe (Fe) 3+ The concentration is 10mg/mL; TA and Fe 3+ The volume ratio of (1): 2, adding NaOH solution for adjusting the pH value, wherein the NaOH solution is 0.1M NaOH solution, and adjusting the pH value to 8.5; caO (CaO) 2 And TA, fe 3+ The reaction is carried out for 2 hours at room temperature; during centrifugation, the product was collected by rotation at 10000rpm for a centrifugation period of 10 min.
In order to dissolve the liposome raw material, in one embodiment of the invention, the solvent in S3 is ethanol and chloroform, the liposome raw material component is egg yolk lecithin, the dosage is 50mg, the dosage of cholesterol is 9.6mg, the dosage of DSPE-PEG2000 is 6.37mg, and CaO 2 -TA-Fe 3+ The concentration of (2) is 10mg/mL, and the concentration of PArg is 15mg/mL; the hydration time of the aqueous PArg solution was 30min and the suspension was centrifuged at 10000rpm for 10min to collect the product.
To achieve the aim of Lipo CaO 2 -TA-Fe 3+ Nanoparticles of PArg are prepared and manufactured step by step in sequence, in one embodiment of the invention the liposomes in S3 encapsulate Lipo CaO 2 -TA-Fe 3+ In the preparation of nanoparticles of PArg, caO is first prepared 2 -TA-Fe 3+ Nanoparticle, synthetic CaO 2 Thereafter, TA-Fe is electrostatically reacted 3+ Self-assembly of coating on CaO 2 Form CaO on the surface of (C) 2 -TA-Fe 3+ Nanoparticles with a particle size of about 45nm, and then passing CaO through the liposomes 2 -TA-Fe 3+ The nano particles and the PArg are respectively wrapped in the hydrophobic layer and the hydrophilic cavity to obtain Lipo CaO 2 -TA-Fe 3+ (PArg) nanoparticles.
To achieve the aim of preparing Lipo CaO 2 -TA-Fe 3+ PArg, in one embodiment of the present inventionThe Lipo CaO 2 -TA-Fe 3+ PArg is used to turn off Ca pump discharge by Fenton catalytic oxidation effect 2+ And opening Ca 2 + Internal flow channel and ion interference therapy to deliver exogenous Ca 2+ Gas NO treatment to release Ca in endoplasmic reticulum 2+ Application of triple beating interaction to preparation of drugs for comprehensively promoting calcium overload treatment of tumors and loading CaO 2 -TA-Fe 3+ And the liposomes of PArg, for use in the treatment of calcium overload and antitumor.
Example 1
Synthesis of CaO by hydrolysis-precipitation process 2 Nanoparticles: the HA solution (10 mL,15 mg/mL) was stirred at room temperature for a minimum of 3h, followed by CaCl 2 (2 mL,0.1 g/mL) solution was added to the HA solution and stirring was continued, followed by the sequential addition of 1mLNH 3 ·H 2 O (1M) and 1.5mL 30% H 2 O 2 And reacting for 3h, adding 0.5mLNaOH (1M) under ultrasonic wave, performing ultrasonic treatment for 30min to obtain white emulsion, washing corresponding HA-stabilized CaO with NaOH (0.1M), water and ethanol 2 The nanoparticles were finally stored in ethanol at 4 ℃ for further use, and the lyophilized CaO was then subjected to a freeze-drying process 2 (2 mg) was dissolved in 2mL of ultra-pure water under ultrasonic treatment, followed by immediately dissolving TA (40 mg/mL, 40. Mu.L) and Fe 3+ (10 mg/mL, 80. Mu.L) was poured into a stirred flask, then NaOH solution (1M) was slowly added to the mixture until the pH of the solution reached 8.5, and finally the resulting product was centrifuged at 10000rpm for 10min to collect the product, which was washed 3 times with ultra pure water, dried in a freeze-dryer, and stored;
CaO produced 2 -TA-Fe 3+ The schematic structure of the nanoparticle is shown in FIG. 1, TA and CaO 2 HA is hydrogen-bonded, TA is hydrogen-bonded with Fe 3+ Are connected by coordination bonds, thereby, in CaO 2 A structure in which a coating film is formed on the surface;
CaO by transmission electron microscopy 2 -TA-Fe 3+ The morphology of the nano particles is characterized, and the sample preparation method comprises the following steps: after the sample was dispersed to a reasonable concentration with water, small drops of the solution were taken and added dropwise to a copper mesh and dried overnight. The results are shown in FIG. 2, and CaO is evident from FIG. 2 2 -TA-Fe 3+ The nanometer particles have uniform morphology and the particle diameter is about 45nm.
CaO analysis by dynamic light scattering and laser particle sizer 2 -TA-Fe 3+ And CaO (CaO) 2 As can be seen from FIG. 3, the particle diameter and the potential size of the nanoparticle are determined by TA-Fe 3+ Coating on CaO 2 Film formation around the surface, caO 2 And CaO (CaO) 2 -TA-Fe 3+ The diameter of the nanoparticle increases from 30nm to 45nm. At the same time, caO 2 Is 18.5mV, while CaO 2 -TA-Fe 3+ Has a zeta potential of 1.58mV, indicating CaO 2 -TA-Fe 3+ The nanoparticle preparation was successful.
Example 2
For the preparation of Lipo CaO 2 -TA-Fe 3+ Palg, egg yolk lecithin (50 mg), cholesterol (9.6 mg) and DSPE-PEG2000 (6.37 mg) were first dissolved in 6mL chloroform in a glass flask, and 1mL CaO was added 2 -TA-Fe 3+ Ethanol (10 mg/mL) was added to the mixture. After 1h of rotary evaporation, a thin lipid film was formed. 5mL Arg (15 mg/mL) was added to the lipid membrane, and the mixture was subjected to continuous ultrasonic treatment at room temperature for 10 minutes and ultrasonic treatment at 0℃for 3 minutes (220W) to obtain a uniform lipid suspension. The suspension was centrifuged at 10000rpm for 10 minutes and washed 3 times with ultra pure water to remove unencapsulated pag. A control group of liposomes was prepared following a similar procedure without the addition of pag; finally, the liposomes were dispersed in PBS buffer.
The obtained Lipo CaO 2 -TA-Fe 3+ The electron microscopy and particle size distribution of the/PArg nanoparticle are shown in FIG. 4, and it can be seen from FIG. 4 that Lipo CaO 2 -TA-Fe 3+ The particle size of the PArg nanoparticle is about 200 nm.
Example 3
Lipo CaO 2 -TA-Fe 3+ PArg nanoparticle and Lipo CaO 2 -TA-Fe 3+ 、Lipo CaO 2 Toxicity data graph for 4T1 cells;
breast cancer cells 4T1 at 5 x 10 3 Inoculating each well into 96-well cell culture plate with volume of 200uL per well, culturing for 24 hr, and adding different concentrations of different test samplesAnd processing: (i) Control; (ii) Lipo CaO 2 ；(iii)Lipo CaO 2 -TA-Fe 3+ ；(ⅳ)Lipo CaO 2 -TA-Fe 3+ Perg, culturing for 48 hours, removing old culture medium, and adding fresh culture medium containing 10 mu LCCK-8 solution per 100uL of each well; after incubation for 1h, the 96-well plate was subjected to Optical Density (OD) measurement at 450nm, the experimental results are shown in FIG. 5, the concentrations of the respective samples are 8ug/mL, 16 ug/mL, 32 ug/mL, 64 ug/mL, 128 ug/mL, 256 ug/mL, 512 ug/mL and 1024 ug/mL, respectively, and the experimental results show that the concentrations are 8ug/mL, 16 ug/mL, 32 ug/mL, 64 ug/mL, 128 ug/mL, 256 ug/mL, 512 ug/mL, and 1024 ug/mL, respectively, relative to Lipo CaO 2 -TA-Fe 3+ And Lipo CaO 2 After treatment, lipo CaO 2 -TA-Fe 3+ The increase of the PArg along with the concentration shows better effect of inhibiting proliferation of breast cancer cells 4T1, and the survival rate of tumor cells is obviously reduced.
Example 4
Lipo CaO 2 -TA-Fe 3+ The PArg can promote apoptosis of breast cancer cells 4T 1;
breast cancer cells 4T1 at 5 x 10 5 Inoculating each hole into a 6-hole cell culture plate, wherein the volume of each hole of culture medium is 3mL, and culturing for 24 hours; adding a different process (i) Control; (ii) Lipo CaO 2 ；(iii)Lipo CaO 2 -TA-Fe 3+ ；(ⅳ)Lipo CaO 2 -TA-Fe 3+ Respectively culturing for 24 hours; firstly, collecting the supernatant into a centrifuge tube, carefully digesting and collecting a cell culture solution into the centrifuge tube by using pancreatin, centrifuging about 500g for 5 minutes, and precipitating cells; washing the cells twice with precooled PBS, centrifuging about 500g for 5 minutes, and collecting the cells; 100uL of pre-chilled 1*AnnexinV Binding Buffer was added to resuspend cells; 5uL of annexin V-FITC and 5uL of PI are added, mixed gently and reacted for 15 minutes at room temperature in a dark place; 400 μl of pre-cooled 1*AnnexinV Binding Buffer was added, mixed gently, and the sample was placed on ice in a dark place, detected by flow cytometry within 1h, and the detection results were analyzed.
As shown in FIG. 6, it was found that Lipo CaO was obtained by flow cytometry analysis 2 -TA-Fe 3+ The PArg can obviously promote the apoptosis of breast cancer cells 4T1, has obvious difference compared with a control group, and has good effect of inhibiting the growth of tumors.
Example 5
Lipo CaO 2 -TA-Fe 3+ Pharmacodynamic research experiments of the PArg nano particles;
female Balb/c mice with age of 6 weeks are purchased, breast cancer cells of 4T1 mice are inoculated subcutaneously on the back of the right upper limbs of the mice, tumor volume is measured after 7 days, and the tumor volume is not less than 100mm 3 Mice with similar tumor volumes and weights were randomly divided into 4 groups of 5 mice each, and the specific groupings were as follows: physiological saline group, lipo CaO 2 Group, lipo CaO 2 -TA-Fe 3+ Group and include Lipo CaO 2 -TA-Fe 3+ group/PArg; the injection dose is 1.5mg/kg, the injection dose is 100 microliters, the administration mode of each group of mice adopts tail intravenous injection, the mice are administrated once every two days for 7 times, the daily normal diet of the mice is ensured in the whole experimental process, the weight of each mouse is weighed every two days, the long diameter (A) and the short diameter (B) of the sarcoma of the tumor-bearing mice are measured by using a digital vernier caliper, and the tumor volume V=A.times.B is calculated according to the formula 2 2, calculating tumor volume; the results are shown in FIG. 7, which shows Lipo CaO 2 -TA-Fe 3+ The pharmaceutical effect of the PArg group is obvious, and the growth of tumor can be greatly inhibited.
The above description is only of the preferred embodiments of the present invention and is not intended to limit the present invention, and various modifications and variations may be made to the present invention by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. Multi-channel Ca for promoting tumor calcium overload 2+ The preparation method of the nano regulator is characterized by comprising the following steps:
s1, synthesizing CaO through hydrolysis-precipitation process 2 Nanoparticles: adding CaCl into sodium hyaluronate solution 2 Adding ammonia water and 30% hydrogen peroxide solution in turn, adding sodium hydroxide aqueous solution under ultrasonic wave after stirring, washing with NaOH, water and ethanol, and freeze-drying for further use;
S2、preparation of CaO 2 -TA-Fe 3+ Nanoparticles: caO prepared in S1 2 Dissolving the nano particles, and then sequentially adding TA aqueous solution and FeCl 3 Adding NaOH solution into the aqueous solution to adjust the pH value, centrifugally collecting the product, washing the product with ultrapure water for a plurality of times, and drying the product in a freeze dryer to obtain CaO 2 -TA-Fe 3+ A nanoparticle;
s3, preparation of LipoCaO 2 -TA-Fe 3+ (pag) nanoparticles: caO prepared in S2 2 -TA-Fe 3+ The nanoparticles were dissolved in ethanol, then dissolved in a glass flask together with the liposome raw material, film was formed by rotary evaporation, then an aqueous polyarginine solution was added to the flask under ultrasound, then sonicated at 0 ℃ for 3 minutes to obtain a uniform lipid suspension, the suspension was centrifuged for 10 minutes, washed three times with ultra pure water to remove unencapsulated pagg, and finally the liposomes were dispersed in PBS buffer.
2. A multi-channel tumorigenic calcium-overloaded Ca according to claim 1 2+ A method for preparing a nano regulator is characterized in that the LipoCaO in S3 2 -TA-Fe 3+ The PArg nanoparticle takes lecithin liposome as a drug carrier, and calcium peroxide composite particles and polyarginine as a hydrophobic layer and a hydrophilic cavity substance.
3. A multi-channel tumorigenic calcium-overloaded Ca according to claim 2 2+ The preparation method of the nano regulator is characterized in that the calcium peroxide composite particles are calcium peroxide-tannic acid-iron and have a chemical formula of CaO 2 -TA-Fe 3+ The polyarginine is poly-L-arginine, namely PArg.
4. A multi-channel tumorigenic calcium-overloaded Ca according to claim 1 2+ The preparation method of the nano regulator is characterized in that the LipoCaO 2 -TA-Fe 3+ The preparation of the/PArg nanoparticles is carried out by reacting CaO 2 -TA-Fe 3+ And lecithin, cholesterol, DSPE-PEG2000 in organic solution by rotary evaporation to form filmAnd is obtained by the hydration of PArg.
5. A multi-channel tumorigenic calcium-overloaded Ca according to claim 1 2+ The preparation method of the nano regulator is characterized in that the sodium hyaluronate solvent in S1 is 10mL,15mg/mL, stirring is required to be carried out for more than 3 hours at room temperature, and CaCl is adopted 2 The aqueous solution was 0.1g/mL, and NH 3 ·H 2 O is 1M, which is prepared in the prior art, and 30% hydrogen peroxide solution is H with the mass fraction of 30% 2 O 2 The stirring reaction time after the addition of the aqueous solution was 3 hours.
6. A multi-channel tumorigenic calcium-overloaded Ca according to claim 1 2+ The preparation method of the nano regulator is characterized in that the aqueous solution of NaOH in the S1 is 0.1M, and the washing process of NaOH, water and ethanol is that NaOH is firstly used for washing treatment to remove superfluous acid, water is used for washing to remove the neutralized solution, then ethanol is used for washing, and the residual water stain is washed and removed.
7. A multi-channel tumorigenic calcium-overloaded Ca according to claim 1 2+ The preparation method of the nano regulator is characterized in that the TA concentration in the S2 is 40mg/mL; fe (Fe) 3+ The concentration is 10mg/mL; TA and Fe 3+ The volume ratio of (1): 2, adding NaOH solution for adjusting the pH value, wherein the NaOH solution is 0.1M NaOH solution, and adjusting the pH value to 8.5; caO (CaO) 2 And TA, fe 3+ The reaction is carried out for 2 hours at room temperature; during centrifugation, the product was collected by rotation at 10000rpm for a centrifugation period of 10 min.
8. A multi-channel tumorigenic calcium-overloaded Ca according to claim 1 2+ The preparation method of the nano regulator is characterized in that the solvent in the S3 is ethanol and chloroform, the liposome raw material component is egg yolk lecithin, the dosage is 50mg, the dosage of cholesterol is 9.6mg, the dosage of DSPE-PEG2000 is 6.37mg, andCaO 2 -TA-Fe 3+ the concentration of (2) is 10mg/mL, and the concentration of PArg is 15mg/mL; the hydration time of the aqueous PArg solution was 30min and the suspension was centrifuged at 10000rpm for 10min to collect the product.
9. A multi-channel tumorigenic calcium-overloaded Ca according to claim 1 2+ The preparation method of the nano regulator is characterized in that the LipoCaO is wrapped by the liposome in the S3 2 -TA-Fe 3+ In the preparation of nanoparticles of PArg, caO is first prepared 2 -TA-Fe 3+ Nanoparticle, synthetic CaO 2 Thereafter, TA-Fe is electrostatically reacted 3+ Self-assembly of coating on CaO 2 Form CaO on the surface of (C) 2 -TA-Fe 3+ Nanoparticles with a particle size of about 45nm, and then passing CaO through the liposomes 2 -TA-Fe 3+ The nano particles and the PArg are respectively wrapped in the hydrophobic layer and the hydrophilic cavity to obtain LipoCaO 2 -TA-Fe 3+ (PArg) nanoparticles.
10. A multi-channel tumorigenic calcium-overloaded Ca according to claim 1 2+ The preparation method of the nano regulator is characterized in that the LipoCaO 2 -TA-Fe 3+ PArg is used to turn off Ca pump discharge by Fenton catalytic oxidation effect 2+ And opening Ca 2+ Internal flow channel and ion interference therapy to deliver exogenous Ca 2+ Gas NO treatment to release Ca in endoplasmic reticulum 2+ Application of triple beating interaction to preparation of drugs for comprehensively promoting calcium overload treatment of tumors and loading CaO 2 -TA-Fe 3+ And the liposomes of PArg, for use in the treatment of calcium overload and antitumor.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310036155.XA CN116440078A (en) | 2023-01-08 | 2023-01-08 | Multi-channel Ca for promoting tumor calcium overload 2+ Preparation method of nano regulator |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310036155.XA CN116440078A (en) | 2023-01-08 | 2023-01-08 | Multi-channel Ca for promoting tumor calcium overload 2+ Preparation method of nano regulator |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN116440078A true CN116440078A (en) | 2023-07-18 |
Family
ID=87130955
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310036155.XA Pending CN116440078A (en) | 2023-01-08 | 2023-01-08 | Multi-channel Ca for promoting tumor calcium overload 2+ Preparation method of nano regulator |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116440078A (en) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20140065246A1 (en) * | 2003-05-30 | 2014-03-06 | Charles E. Zeilig | Intracellular calcium modulation for cancer treatment |
| CN110179818A (en) * | 2019-05-27 | 2019-08-30 | 河北大学 | CaO2@DOX@ZIF-67 nano material and the preparation method and application thereof |
| US20200069727A1 (en) * | 2017-05-04 | 2020-03-05 | University Of Ulster | Calcium peroxides nanoparticles as adjuvant therapy |
| CN112274495A (en) * | 2020-10-31 | 2021-01-29 | 郑州大学 | H2O2Preparation method and application of self-supply type calcium peroxide loaded curcumin nanoparticles |
-
2023
- 2023-01-08 CN CN202310036155.XA patent/CN116440078A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20140065246A1 (en) * | 2003-05-30 | 2014-03-06 | Charles E. Zeilig | Intracellular calcium modulation for cancer treatment |
| US20200069727A1 (en) * | 2017-05-04 | 2020-03-05 | University Of Ulster | Calcium peroxides nanoparticles as adjuvant therapy |
| CN110179818A (en) * | 2019-05-27 | 2019-08-30 | 河北大学 | CaO2@DOX@ZIF-67 nano material and the preparation method and application thereof |
| CN112274495A (en) * | 2020-10-31 | 2021-01-29 | 郑州大学 | H2O2Preparation method and application of self-supply type calcium peroxide loaded curcumin nanoparticles |
Non-Patent Citations (2)
| Title |
|---|
| FEI CHEN等: ""A CaO2@Tannic Acid-FeIII Nanoconjugate for Enhanced Chemodynamic Tumor Therapy"", 《CHEMMEDCHEM》, no. 16, 31 December 2021 (2021-12-31), pages 2278 * |
| JUNJIE LIU: ""Boosting tumor treatment by dredging the hurdles of chemodynamic therapy synergistic ion therapy"", 《CHEMICAL ENGINEERING JOURNAL》, no. 411, 9 January 2021 (2021-01-09), pages 128440 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US11918686B2 (en) | Lipid bilayer coated mesoporous silica nanoparticles with a high loading capacity for one or more anticancer agents | |
| KR101720851B1 (en) | Nanovesicles derived from cell membrane and use thereof | |
| Alshaer et al. | Encapsulation of echinomycin in cyclodextrin inclusion complexes into liposomes: in vitro anti-proliferative and anti-invasive activity in glioblastoma | |
| He et al. | Enhancement of cisplatin efficacy by lipid–CaO 2 nanocarrier-mediated comprehensive modulation of the tumor microenvironment | |
| CN113355290B (en) | Anti-tumor engineered exosome, preparation method and application | |
| Yu et al. | Near-infrared photoactivatable semiconducting polymer nanocomplexes with bispecific metabolism interventions for enhanced cancer immunotherapy | |
| Zhang et al. | A smart O2-generating nanocarrier optimizes drug transportation comprehensively for chemotherapy improving | |
| Dai et al. | Nanomedicines modulating myeloid-derived suppressor cells for improving cancer immunotherapy | |
| He et al. | pH-Responsive size-shrinkable mesoporous silica-based nanocarriers for improving tumor penetration and therapeutic efficacy | |
| Shen et al. | Lipid-polymer hybrid nanoparticle with cell-distinct drug release for treatment of stemness-derived resistant tumor | |
| CN109953974B (en) | Preparation method of enzyme-reduction dual-responsiveness hyaluronic acid-polypropylene sulfide copolymer nanocapsule | |
| Li et al. | A hybrid of lactic acid bacteria and metal-organic frameworks potentiates photodynamic immunotherapy for hypoxia-targeted tumor eradication | |
| CN116440078A (en) | Multi-channel Ca for promoting tumor calcium overload 2+ Preparation method of nano regulator | |
| CN116251062A (en) | Preparation method and application of bacterial membrane-liposome drug-loading system | |
| CN108888764B (en) | Nano drug delivery system based on low-generation PAMAM dendrimer loaded disulfiram and photosensitizer indocyanine green and application thereof | |
| CN110859820A (en) | Biocompatible alkaline earth metal peroxide nano preparation, preparation method and application thereof | |
| CN112274495B (en) | H2O2Preparation method and application of self-supply type calcium peroxide loaded curcumin nanoparticles | |
| CN111840254B (en) | Nano realgar compound medicine and its preparing method and use | |
| CN114681430A (en) | Resveratrol lecithin nanoparticles and preparation method and application thereof | |
| CN113679851A (en) | Enzyme response hybrid nanoparticle coated with Ganisertib and tadalafil and preparation method and application thereof | |
| Imanimoghadam et al. | Improving Chemotherapy Effectiveness: Utilizing CuS Nanoparticles Coated with AS1411 Aptamer and Chitosan for Targeted Delivery of Doxorubicin to Cancerous Cells | |
| Wang et al. | Local delivery of artesunate dimer liposomes incorporated injectable hydrogel for H2O2 and pH-independent chemodynamic therapy | |
| Sudareva et al. | Comparison of Delivery Systems for Chemotherapy Preparation Doxorubicin using Electron Microscopic and Hydrodynamic Methods | |
| CN116603069B (en) | Microorganism targeting implantation system and method | |
| CN115120736B (en) | Multifunctional gene vector and application thereof in delivery of miRNA |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |